Investigating the Role of E3 Ubiquitin Ligases in Flavivirus Infection

黄病毒，如登革热（DENV）和寨卡病毒（ZIKV）构成了严重的公共卫生挑战，每年感染全球超过4亿人。尽管如此，目前还没有有效的疫苗或抗病毒药物来对抗黄病毒，这突出了我们需要提高对其生物学的理解。虽然已知黄病毒劫持宿主泛素化系统以发挥其优势，但催化泛素化的E3连接酶在黄病毒生命周期中的作用受到有限的关注，对正常E3连接酶生物学的理解有限。该项目旨在鉴定和鉴定宿主细胞中的E3-连接酶，这些酶被增选对黄病毒生命周期具有前病毒作用。我们将应用无偏的以及基于候选人的方法来鉴定在ZIKV感染中具有改变的活性的E3连接酶。我们将结合联合收割机的遗传，生物化学和蛋白质组学策略，进行详细的表征这些E3-连接酶在病毒复制。最后，我们的目标是验证这些E3-连接酶介导的过程在蚊子载体中的保守性，以确定在人类和载体宿主中的普遍感染机制。从这项研究中获得的见解将进一步加深我们对以下问题的理解：健康的E3-连接酶生物学和人类的免疫功能，黄病毒生物学，并可能有助于开发广谱抗病毒药物。BBSRC相关领域这项工作属于更广泛的BBSRC主题“生物科学对健康的综合理解”，特别是优先领域“终生健康老龄化”和“终身健康和福祉”。我们将获得E3-连接酶细胞功能的基本方面的理解，黄病毒感染使我们能够确定E3-连接酶的关键相互作用和活动，这对调节更广泛的细胞生物学至关重要。鉴定在不同黄病毒物种和不同宿主物种中具有保守功能的E3连接酶也将使我们能够鉴定核心E3连接酶，其在细胞过程如代谢和免疫应答中是关键的。总之，这将提高我们对细胞生物学的理解，并确定关键的E3-连接酶，这些酶对细胞的健康功能很重要，特别是对疾病中的功能障碍很敏感。

Flaviviruses such as Dengue (DENV) and Zika (ZIKV) pose serious public health challenges, infecting over 400 million people a year globally. Despite this, no effective vaccines or antivirals are currently available to combat flaviviruses, highlighting the need to improve our understanding of their biology. While it is known that flaviviruses hijack the host ubiquitylation system to their advantage, E3-ligases that catalyse ubiquitylation have received limited attention on their role in the flavivirus lifecycle, with limited understanding of normal E3-ligase biology. This project aims to identify and characterise E3-ligases of the host cell that are co-opted to have a proviral effect on the flavivirus lifecycle. We will apply both an unbiased as well as a candidate-based approach to identify E3-ligases with altered activity in ZIKV infection. We will combine genetic, biochemical and proteomics strategies to perform detailed characterization of these E3- ligases in virus replication. Finally, we aim to verify the conservation of these E3-ligase-mediated processes in the mosquito vector to define universal infection mechanisms in both human and the vector host. The insights gained from this study will further our understanding of; healthy E3-ligase biology and immune function in humans, flavivirus biology, and potentially contribute to the development of broad-spectrum antivirals.BBSRC Relevant Areas This work comes under the wider BBSRC theme of 'Bioscience for an integrated understanding of health', specifically the priority areas 'Healthy ageing across the life course' and 'Lifelong health and wellbeing'. We will gain an understanding of the fundamental aspects of E3-ligase cellular function, with flavivirus infection allowing us to identify key interactions and activities of E3-ligases which are crucial to modulating wider cellular biology. Identifying E3-ligases with conserved functions across different flavivirus species and in different host species will also allow us to identify core E3-ligases which are key in cellular processes such as metabolism and the immune response. Together, this will improve our understanding of cellular biology and also identify key E3-ligases which are important in healthy function of the cell and particularly susceptible to dysfunction in disease.